Cloning and functional analysis of the Ldcht10 gene sequence in Lymantria dispar larvae
Author of the article:LIU Jian-Hong** ZHANG Chang ZHAO Qiu-Yong FAN Xiao-Jun***
Author's Workplace：Department of Biological and Pharmaceutical Engineering, College of Chemical Engineering and Technology, Taiyuan University of Technology, Taiyuan 030024, China
Key Words：Lymantria dispar, chitinases, qRT-PCR, RNAi
Abstract： [Objectives] In order to provide a basis for effective pest control based on RNAi, the temporal and spatial expression, and the biological functions of the chitinase family of genes in Lymantria dispar were investigated and chitinase genes causing death during development were screened. [Methods] Degenerate primers were designed to clone the critical sequence of LdCht10 and the real-time quantitative PCR method was used to detect the relative expression level of LdCht10 in different instars and tissues of Lymantria dispar larvae. A fragment of the cloned sequence was chosen to study gene function using the RNA interference method. [Results] A critical sequence of LdCht10 of 2 057 bp in length was successfully cloned. The RT-PCR results show that the temporal and spatial expression patterns of LdCht10 were significantly different, and that the LdCht10 gene was expressed in all instars and tissues, with the highest expression occurring in the foregut and hindgut. RNA interference showed that 24.3% of test larvae died as a result of not completing molting and pupation after the gene was silenced. [Conclusion] The temporal and spatial expression profiles of LdCht10 were distinct in different instars and tissues. Based on the RNAi results, LdCht10 could play an important role in the molting process of L. dispar, and gene silencing can block ecdysis causing death in some larvae.